Nitric oxide synthase expression and cell changes in dorsal root ganglia and spinal dorsal horn of the developing and adult Rana esculenta implicate a role of nitric oxide in limb metamorphosis

Metamorphosis of amphibians requires reconfiguration of sensory and locomotor neural networks. In view of such plastic changes and implications of nitric oxide (NO) in neural developmental shaping, we examined via histochemistry and immunohistochemistry its synthetic enzyme nitric oxide synthase (NOS) in dorsal root ganglia (DRGs) and dorsal horn of the developing and adult frog Rana esculenta. In limb DRGs, NOS positivity was first and selectively detected just before limb bud appearance, increased during metamorphosis, and was then down-regulated. In adulthood, NOS was expressed in some DRG neurons at all segmental levels. Similar features were detected in the dorsal horn neuropil. In limb DRGs, cell counts in Nissl-stained sections revealed a twofold increase of differentiated neurons during metamorphosis and an additional twofold increase in adulthood. Perikaryal sizes in limb DRGs did not vary during metamorphosis but increased and were more heterogeneous in the adult frog, probably reflecting adaptation to body size. NOS and cell changes during metamorphosis were much less marked in DRGs at other levels. Carbocyanine tracing documented selective labeling of NOS-expressing hindlimb DRG neurons from the spinal nerve at the time of initiation of hindlimb movements. The findings show that, in limb DRG neurons, NOS parallels cell differentiation and limb development during metamorphosis. The data also provide evidence of NOS expression in DRG cells innervating the hindlimbs when sensorimotor circuits become functionally mature. This study indicates a key role of NO production in the maturation of sensory functions that subserves in amphibians the transition from swimming to tetrapod locomotion. © 2004 Wiley-Liss, Inc.